C3, C4, and the terminal complement complex differ from C1q by binding predominantly to the antigenic part of solid phase immune complexes.
The binding of the C components C1q, C4, C3, the terminal C5b-9 complement complex (TCC) and S protein to immune complexes was studied. The hapten 5-iodo-4-hydroxy-3-nitrophenacetyl (NIP) conjugated to BSA was adsorbed to polystyrene plates and reacted with a human IgG3-mouse chimeric anti-NIP antibody. After addition of serum a dose-dependent binding of C1q, C4, C3, and TCC to the immune complexes was found. An increase in the amount of NIP-BSA was associated with an increase in the binding of TCC and a decrease in the binding of S-protein. After addition of soluble NIP only 4 to 6% of the anti-NIP antibody remained bound to the Ag. C1q showed diminished binding after addition of NIP, whereas C4, C3, and TCC quantitatively remained bound to the Ag. Binding of TCC to the immune complexes was also found in an alternative assay, in which the anti-NIP antibody was adsorbed to the solid phase before NIP-BSA and an additional layer of anti-NIP antibody were added. The supernatants from the solid phase assay were tested for C3 activation and formation of the fluid phase TCC (SC5b-9). Activation of the C3 was reflected in the fluid phase by a dose-dependent increase in C3 activation products. This was not seen for TCC despite increased binding to the solid phase.
Garred
P
P
Institute of Immunology and Rheumatology, National Hospital, University of Oslo, Norway.
Michaelsen
T E
TE
Aase
A
A
Mollnes
T E
TE
D000818:Animals; D000936:Antigen-Antibody Complex / Q000378:metabolism*; D001665:Binding Sites; D015922:Complement C1q / Q000378:metabolism*; D003176:Complement C3 / Q000378:metabolism*; D003181:Complement C4 / Q000378:metabolism*; D015938:Complement Membrane Attack Complex / Q000378:metabolism*; D007074:Immunoglobulin G / Q000378:metabolism; D066298:In Vitro Techniques; D051379:Mice; D012995:Solubility
D000936:Antigen-Antibody Complex; D003176:Complement C3; D003181:Complement C4; D015938:Complement Membrane Attack Complex; D007074:Immunoglobulin G; D015922:Complement C1q